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goldengate cloning site  (GoldenGate Software Inc)

 
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    GoldenGate Software Inc goldengate cloning site
    Cas9 and Cas12a toolkit parts available via AddGene. Plasmid parts of the toolkit: Level 1 guide accepters for Cas9 and Cas12a with three promoter options. Compatible binary vectors containing plant selection markers and nuclease cassettes which are wheat or barley specific. P- 35S = Cauliflower mosaic virus long 35S promoter, P- Pv Ubi = Panicum virgatum ubiquitin promoter, HptII = hygromycin phosphotransferase CDS with intron, T- 35S = Cauliflower mosaic virus terminator, P- Zm Ubi = Zea mays ubiquitin promoter, tt At Cas12a + 8int = temperature tolerant Arabidopsis codon optimised Cas12a CDS with 8 introns, Zm Cas9 + 13int = Zea mays codon optimised Cas9 with thirteen introns, T-Nos = nopaline synthase terminator, GRF-GIF = Growth-regulating factor 4 (GRF4) and its cofactor GRF interacting factor 1(GRF1) fusion, GR-GRF-GIF = Rat glucocorticoid receptor (GR) & Growth-regulating factor 4 (GRF4) & its cofactor GRF interacting factor 1(GRF1) fusion. RFP = red fluorescent protein for colour selection during <t>cloning,</t> Ta U6 = Triticum aestivum U6 promoter, Ta U3 = Triticum aestivum U3 promoter, Hv U3 = Hordeum vulgare U3 promoter, HH = Hammerhead ribozyme, HDV = hepatitis delta virus ribozyme, tRNA = tRNA sequence, DR = short invariable 5’ region of Cas12a guide, sgRNA = invariable 3’ region of Cas9 guide, LacZ = blue/white colour selection marker for cloning, BsaI = restriction <t>site,</t> BpiI = restriction site. Triangles indicate GoldenGate cloning site <t>cut</t> <t>points.</t> Arrows indicate direction of transcription
    Goldengate Cloning Site, supplied by GoldenGate Software Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/goldengate+cloning+sites/goldengate+cloning+site/pmc11321142-240-3-2
    Average 90 stars, based on 1 article reviews
    goldengate cloning site - by Bioz Stars, 2026-10
    90/100 stars

    Images

    1) Product Images from "An optimised CRISPR Cas9 and Cas12a mutagenesis toolkit for Barley and Wheat"

    Article Title: An optimised CRISPR Cas9 and Cas12a mutagenesis toolkit for Barley and Wheat

    Journal: Plant Methods

    doi: 10.1186/s13007-024-01234-y

    Cas9 and Cas12a toolkit parts available via AddGene. Plasmid parts of the toolkit: Level 1 guide accepters for Cas9 and Cas12a with three promoter options. Compatible binary vectors containing plant selection markers and nuclease cassettes which are wheat or barley specific. P- 35S = Cauliflower mosaic virus long 35S promoter, P- Pv Ubi = Panicum virgatum ubiquitin promoter, HptII = hygromycin phosphotransferase CDS with intron, T- 35S = Cauliflower mosaic virus terminator, P- Zm Ubi = Zea mays ubiquitin promoter, tt At Cas12a + 8int = temperature tolerant Arabidopsis codon optimised Cas12a CDS with 8 introns, Zm Cas9 + 13int = Zea mays codon optimised Cas9 with thirteen introns, T-Nos = nopaline synthase terminator, GRF-GIF = Growth-regulating factor 4 (GRF4) and its cofactor GRF interacting factor 1(GRF1) fusion, GR-GRF-GIF = Rat glucocorticoid receptor (GR) & Growth-regulating factor 4 (GRF4) & its cofactor GRF interacting factor 1(GRF1) fusion. RFP = red fluorescent protein for colour selection during cloning, Ta U6 = Triticum aestivum U6 promoter, Ta U3 = Triticum aestivum U3 promoter, Hv U3 = Hordeum vulgare U3 promoter, HH = Hammerhead ribozyme, HDV = hepatitis delta virus ribozyme, tRNA = tRNA sequence, DR = short invariable 5’ region of Cas12a guide, sgRNA = invariable 3’ region of Cas9 guide, LacZ = blue/white colour selection marker for cloning, BsaI = restriction site, BpiI = restriction site. Triangles indicate GoldenGate cloning site cut points. Arrows indicate direction of transcription
    Figure Legend Snippet: Cas9 and Cas12a toolkit parts available via AddGene. Plasmid parts of the toolkit: Level 1 guide accepters for Cas9 and Cas12a with three promoter options. Compatible binary vectors containing plant selection markers and nuclease cassettes which are wheat or barley specific. P- 35S = Cauliflower mosaic virus long 35S promoter, P- Pv Ubi = Panicum virgatum ubiquitin promoter, HptII = hygromycin phosphotransferase CDS with intron, T- 35S = Cauliflower mosaic virus terminator, P- Zm Ubi = Zea mays ubiquitin promoter, tt At Cas12a + 8int = temperature tolerant Arabidopsis codon optimised Cas12a CDS with 8 introns, Zm Cas9 + 13int = Zea mays codon optimised Cas9 with thirteen introns, T-Nos = nopaline synthase terminator, GRF-GIF = Growth-regulating factor 4 (GRF4) and its cofactor GRF interacting factor 1(GRF1) fusion, GR-GRF-GIF = Rat glucocorticoid receptor (GR) & Growth-regulating factor 4 (GRF4) & its cofactor GRF interacting factor 1(GRF1) fusion. RFP = red fluorescent protein for colour selection during cloning, Ta U6 = Triticum aestivum U6 promoter, Ta U3 = Triticum aestivum U3 promoter, Hv U3 = Hordeum vulgare U3 promoter, HH = Hammerhead ribozyme, HDV = hepatitis delta virus ribozyme, tRNA = tRNA sequence, DR = short invariable 5’ region of Cas12a guide, sgRNA = invariable 3’ region of Cas9 guide, LacZ = blue/white colour selection marker for cloning, BsaI = restriction site, BpiI = restriction site. Triangles indicate GoldenGate cloning site cut points. Arrows indicate direction of transcription

    Techniques Used: Plasmid Preparation, Selection, Virus, Ubiquitin Proteomics, Cloning, Sequencing, Marker

    Toolkit cloning strategy. Toolkit main cloning routes: A = level 1 oligo cloning to give guide cassettes (black boxes with red outline), B = Up to 4 level 1 guides directly cloned into level 2 accepter, C = more than 4 guides cloned into level 2 accepter via intermediate level M step. Thin arrows represent complementary oligonucleotides that will later transcribe as the protospacer. Thick arrows show the direction of cloning. U6/3 = U6 or U3 promoter (Note – the invariable DR of Cas12a guide and sgRNA of Cas9 guide are not shown in level 1 guide accepters, but they are present outside of the BsaI-BsaI fragment. LacZ = blue/white colour selection marker for cloning, RFP = red fluorescent protein for colour selection during cloning, BsaI = restriction site, BpiI = restriction site. Triangles indicate GoldenGate cloning site cut points, HptII = hygromycin phosphotransferase plant selection marker, Cas = Cas9 or Cas12a nuclease cassette
    Figure Legend Snippet: Toolkit cloning strategy. Toolkit main cloning routes: A = level 1 oligo cloning to give guide cassettes (black boxes with red outline), B = Up to 4 level 1 guides directly cloned into level 2 accepter, C = more than 4 guides cloned into level 2 accepter via intermediate level M step. Thin arrows represent complementary oligonucleotides that will later transcribe as the protospacer. Thick arrows show the direction of cloning. U6/3 = U6 or U3 promoter (Note – the invariable DR of Cas12a guide and sgRNA of Cas9 guide are not shown in level 1 guide accepters, but they are present outside of the BsaI-BsaI fragment. LacZ = blue/white colour selection marker for cloning, RFP = red fluorescent protein for colour selection during cloning, BsaI = restriction site, BpiI = restriction site. Triangles indicate GoldenGate cloning site cut points, HptII = hygromycin phosphotransferase plant selection marker, Cas = Cas9 or Cas12a nuclease cassette

    Techniques Used: Cloning, Clone Assay, Selection, Marker

    Related Articles

    Plasmid Preparation:

    Article Title: Arabidopsis and maize terminator strength is determined by GC content, polyadenylation motifs and cleavage probability.
    Article Snippet: The Plant STARR-seq plasmid pPSt (https://www.addgene.org/ 203590/) used in this study is based on the pPSup plasmid (https:// www.addgene.org/149416/)5 with adaptations to enable the characterization of terminators. .. The base plasmid pPSt contains three consecutive GoldenGate cloning sites. .. To generate the terminator libraries, Nature Communications | (2024) 15:5868 15 we first inserted the 35S promoter together with a 5’ UTR derived from themaize histone H3 gene Zm00001d041672, an ATG start codon, and a 18-nucleotide randombarcode (VNNVNNVNNVNNVNN;V = A, CorG) into the upstream Golden Gate site using the restriction enzyme BbsIHF (NEB).

    Cloning:

    Article Title: Arabidopsis and maize terminator strength is determined by GC content, polyadenylation motifs and cleavage probability.
    Article Snippet: The Plant STARR-seq plasmid pPSt (https://www.addgene.org/ 203590/) used in this study is based on the pPSup plasmid (https:// www.addgene.org/149416/)5 with adaptations to enable the characterization of terminators. .. The base plasmid pPSt contains three consecutive GoldenGate cloning sites. .. To generate the terminator libraries, Nature Communications | (2024) 15:5868 15 we first inserted the 35S promoter together with a 5’ UTR derived from themaize histone H3 gene Zm00001d041672, an ATG start codon, and a 18-nucleotide randombarcode (VNNVNNVNNVNNVNN;V = A, CorG) into the upstream Golden Gate site using the restriction enzyme BbsIHF (NEB).



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    Cas9 and Cas12a toolkit parts available via AddGene. Plasmid parts of the toolkit: Level 1 guide accepters for Cas9 and Cas12a with three promoter options. Compatible binary vectors containing plant selection markers and nuclease cassettes which are wheat or barley specific. P- 35S = Cauliflower mosaic virus long 35S promoter, P- Pv Ubi = Panicum virgatum ubiquitin promoter, HptII = hygromycin phosphotransferase CDS with intron, T- 35S = Cauliflower mosaic virus terminator, P- Zm Ubi = Zea mays ubiquitin promoter, tt At Cas12a + 8int = temperature tolerant Arabidopsis codon optimised Cas12a CDS with 8 introns, Zm Cas9 + 13int = Zea mays codon optimised Cas9 with thirteen introns, T-Nos = nopaline synthase terminator, GRF-GIF = Growth-regulating factor 4 (GRF4) and its cofactor GRF interacting factor 1(GRF1) fusion, GR-GRF-GIF = Rat glucocorticoid receptor (GR) & Growth-regulating factor 4 (GRF4) & its cofactor GRF interacting factor 1(GRF1) fusion. RFP = red fluorescent protein for colour selection during <t>cloning,</t> Ta U6 = Triticum aestivum U6 promoter, Ta U3 = Triticum aestivum U3 promoter, Hv U3 = Hordeum vulgare U3 promoter, HH = Hammerhead ribozyme, HDV = hepatitis delta virus ribozyme, tRNA = tRNA sequence, DR = short invariable 5’ region of Cas12a guide, sgRNA = invariable 3’ region of Cas9 guide, LacZ = blue/white colour selection marker for cloning, BsaI = restriction <t>site,</t> BpiI = restriction site. Triangles indicate GoldenGate cloning site <t>cut</t> <t>points.</t> Arrows indicate direction of transcription
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    Cas9 and Cas12a toolkit parts available via AddGene. Plasmid parts of the toolkit: Level 1 guide accepters for Cas9 and Cas12a with three promoter options. Compatible binary vectors containing plant selection markers and nuclease cassettes which are wheat or barley specific. P- 35S = Cauliflower mosaic virus long 35S promoter, P- Pv Ubi = Panicum virgatum ubiquitin promoter, HptII = hygromycin phosphotransferase CDS with intron, T- 35S = Cauliflower mosaic virus terminator, P- Zm Ubi = Zea mays ubiquitin promoter, tt At Cas12a + 8int = temperature tolerant Arabidopsis codon optimised Cas12a CDS with 8 introns, Zm Cas9 + 13int = Zea mays codon optimised Cas9 with thirteen introns, T-Nos = nopaline synthase terminator, GRF-GIF = Growth-regulating factor 4 (GRF4) and its cofactor GRF interacting factor 1(GRF1) fusion, GR-GRF-GIF = Rat glucocorticoid receptor (GR) & Growth-regulating factor 4 (GRF4) & its cofactor GRF interacting factor 1(GRF1) fusion. RFP = red fluorescent protein for colour selection during cloning, Ta U6 = Triticum aestivum U6 promoter, Ta U3 = Triticum aestivum U3 promoter, Hv U3 = Hordeum vulgare U3 promoter, HH = Hammerhead ribozyme, HDV = hepatitis delta virus ribozyme, tRNA = tRNA sequence, DR = short invariable 5’ region of Cas12a guide, sgRNA = invariable 3’ region of Cas9 guide, LacZ = blue/white colour selection marker for cloning, BsaI = restriction site, BpiI = restriction site. Triangles indicate GoldenGate cloning site cut points. Arrows indicate direction of transcription

    Journal: Plant Methods

    Article Title: An optimised CRISPR Cas9 and Cas12a mutagenesis toolkit for Barley and Wheat

    doi: 10.1186/s13007-024-01234-y

    Figure Lengend Snippet: Cas9 and Cas12a toolkit parts available via AddGene. Plasmid parts of the toolkit: Level 1 guide accepters for Cas9 and Cas12a with three promoter options. Compatible binary vectors containing plant selection markers and nuclease cassettes which are wheat or barley specific. P- 35S = Cauliflower mosaic virus long 35S promoter, P- Pv Ubi = Panicum virgatum ubiquitin promoter, HptII = hygromycin phosphotransferase CDS with intron, T- 35S = Cauliflower mosaic virus terminator, P- Zm Ubi = Zea mays ubiquitin promoter, tt At Cas12a + 8int = temperature tolerant Arabidopsis codon optimised Cas12a CDS with 8 introns, Zm Cas9 + 13int = Zea mays codon optimised Cas9 with thirteen introns, T-Nos = nopaline synthase terminator, GRF-GIF = Growth-regulating factor 4 (GRF4) and its cofactor GRF interacting factor 1(GRF1) fusion, GR-GRF-GIF = Rat glucocorticoid receptor (GR) & Growth-regulating factor 4 (GRF4) & its cofactor GRF interacting factor 1(GRF1) fusion. RFP = red fluorescent protein for colour selection during cloning, Ta U6 = Triticum aestivum U6 promoter, Ta U3 = Triticum aestivum U3 promoter, Hv U3 = Hordeum vulgare U3 promoter, HH = Hammerhead ribozyme, HDV = hepatitis delta virus ribozyme, tRNA = tRNA sequence, DR = short invariable 5’ region of Cas12a guide, sgRNA = invariable 3’ region of Cas9 guide, LacZ = blue/white colour selection marker for cloning, BsaI = restriction site, BpiI = restriction site. Triangles indicate GoldenGate cloning site cut points. Arrows indicate direction of transcription

    Article Snippet: Triangles indicate GoldenGate cloning site cut points, HptII = hygromycin phosphotransferase plant selection marker, Cas = Cas9 or Cas12a nuclease cassette Additional file details (in the appropriate tabs) the various level 1, level M, level 2 & end linker plasmids available via AddGene.

    Techniques: Plasmid Preparation, Selection, Virus, Ubiquitin Proteomics, Cloning, Sequencing, Marker

    Toolkit cloning strategy. Toolkit main cloning routes: A = level 1 oligo cloning to give guide cassettes (black boxes with red outline), B = Up to 4 level 1 guides directly cloned into level 2 accepter, C = more than 4 guides cloned into level 2 accepter via intermediate level M step. Thin arrows represent complementary oligonucleotides that will later transcribe as the protospacer. Thick arrows show the direction of cloning. U6/3 = U6 or U3 promoter (Note – the invariable DR of Cas12a guide and sgRNA of Cas9 guide are not shown in level 1 guide accepters, but they are present outside of the BsaI-BsaI fragment. LacZ = blue/white colour selection marker for cloning, RFP = red fluorescent protein for colour selection during cloning, BsaI = restriction site, BpiI = restriction site. Triangles indicate GoldenGate cloning site cut points, HptII = hygromycin phosphotransferase plant selection marker, Cas = Cas9 or Cas12a nuclease cassette

    Journal: Plant Methods

    Article Title: An optimised CRISPR Cas9 and Cas12a mutagenesis toolkit for Barley and Wheat

    doi: 10.1186/s13007-024-01234-y

    Figure Lengend Snippet: Toolkit cloning strategy. Toolkit main cloning routes: A = level 1 oligo cloning to give guide cassettes (black boxes with red outline), B = Up to 4 level 1 guides directly cloned into level 2 accepter, C = more than 4 guides cloned into level 2 accepter via intermediate level M step. Thin arrows represent complementary oligonucleotides that will later transcribe as the protospacer. Thick arrows show the direction of cloning. U6/3 = U6 or U3 promoter (Note – the invariable DR of Cas12a guide and sgRNA of Cas9 guide are not shown in level 1 guide accepters, but they are present outside of the BsaI-BsaI fragment. LacZ = blue/white colour selection marker for cloning, RFP = red fluorescent protein for colour selection during cloning, BsaI = restriction site, BpiI = restriction site. Triangles indicate GoldenGate cloning site cut points, HptII = hygromycin phosphotransferase plant selection marker, Cas = Cas9 or Cas12a nuclease cassette

    Article Snippet: Triangles indicate GoldenGate cloning site cut points, HptII = hygromycin phosphotransferase plant selection marker, Cas = Cas9 or Cas12a nuclease cassette Additional file details (in the appropriate tabs) the various level 1, level M, level 2 & end linker plasmids available via AddGene.

    Techniques: Cloning, Clone Assay, Selection, Marker